Esponjas do gênero Dysidea (Ordem: Dictyoceratida) caracterizam-se por apresentarem grande diversidade de metabólitos secundários, muitos dos quais apresentam potentes atividades biológicas. Este trabalho descreve o estudo de duas amostras de esponjas da espécie Dysidea robusta, DR1 e DR2, coletadas no litoral da Bahia em 1999. Tal estudo consistiu no fracionamento das amostras, nas análises de seus extratos brutos por LC-MS e técnicas de RMN- mono e bidimensionais. Dentre os extratos de DR1, a fração DR1-EP-5A obtida do extrato éter de petróleo apresentou uma mistura de três ceramidas saturadas (22, 23 e 24). Já da amostra DR2, as frações do extrato aquoso DR2-AQ-6B e DR2-AQ-6D mostraram ser constituídas por derivados do ácido pirodisinóico (18, 19, 20 e 21). Com exceção do ácido pirodisinóico (18), os demais compostos isolados ainda não foram relatados na literatura. / Sponges of the genus Dysidea (Order: Dyctioceratida) are characterized as sources of several biologically active secondary metabolites. This work describes the study of two samples of D.robusta, DR1 and DR2, both collected at the Bahia state coastline, in 1999. The investigation aimed the crude extract fractionation and analysis by LC-MS and by 1D and 2D NMR techniques. Among the extracts DR1, the fraction DR1-EP-5A obtained from the petroleum ether extract showed a mixture of three saturated ceramides, represented by 22, 23 and 24. From the DR2 sample, the fractions obtained from the aqueous extract DR2-AQ-6B and -6D presented pirodisinoic acid derivates 18, 19, 20 and 21. Except for pyrodisinoic acid (18), all other isolated compounds haven´t been reported in the literature yet.

Orientador: Alberto José Cavalheiro / Banca: Cíntia Duarte de Freitas Milagre / Banca: Marcelo Telascrêa / Resumo: As 5,6-diidro-2-pironas 6-substituídas, também conhecidas como estirilpironas, são uma importante classe de metabólitos secundários presentes em plantas. São substâncias biologicamente ativas, apresentando como, por exemplo, atividade anticâncer, antioxidante, antifúngica e antiviral. O gênero Cryptocarya, pertence à família Lauraceae e apresenta diversas estirilpironas em suas mais diversas partes: folhas, cascas, sementes e raízes. Foram selecionadas para análise as espécies C. mandioccana, C. moschata e C. botelhensis. Foram utilizadas apenas as folhas, por apresentarem facilidade para coleta e abundância. Este trabalho tem como objetivo a identificação das estirilpironas presentes em três espécies brasileiras de Cryptocarya com o uso das técnicas HPLC-DAD-MS. Foi desenvolvido método cromatográfico para análise do extrato hidroalcoólico das folhas de espécies de Cryptocarya por HPLC-DAD-MS utilizando etanol como fase orgânica na fase móvel. Com os espectros no UV foi possível identificar que as classes de metabólitos presentes nas amostras eram alcaloides, flavonoides e estirilpironas. Utilizando os dados de MS e MS/MS, foi possível a caracterização de estirilpironas presentes nos extratos, assim como a sugestão de suas fragmentações por espectrometria de massas, além da identificação dos íons m/z 163 e m/z 189 característicos da fragmentação das estirilpironas. Foi detectado o íon m/z 423 no extrato de C. moschata, referente a um possível dímero da goniotalamina, sem rela... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The 6 - substituted 5,6 - dihydro - 2 - pyrones, also known as styryl py rone s, are an important class of secondary metabolites present in plants. They are biologically active substances, presenting, for examp le, anticancer, antioxidant, antifungal and antiviral activity. The genus Cryptocarya belongs to the family Lauraceae and presents several styrylpyrones i n its most diverse parts: leaves, barks, seeds and roots. The species C. mandioccana, C. moschata and C. botelhensis were selected for an alysis. We chose to use only leaves, because they are easy to collect and abu n dant . The objective of this work is to identify the styrylpyrones present in three Brazilian Cryptocarya species using the HPLC - DAD - MS technique s. A chromatographic method was developed to analyze the hydroal coholic extract from leaves Cryptocarya species by HPLC - DAD - MS using ethanol as the organic solvent in the mobile phase. With the UV spectra were possible to identify the classes of metabolites present in the samples as alkaloids, flavonoids and styrylpyrones . Addtitionaly, with the MS and MS 2 data, was possible to characterize the styrylpyrones present in the extracts, as well as the suggestion of their fragments by mass spectrometry, in additi on to the identification of íons m/z 163 and m/z 189 characteristics of the f ragmentation of styry lpi rones. The m/ z 423 ion detected in C. moschata extract was tentatively attributed to a goniotalamine dimer, with no previuous reports for Cryptocarya species. W ith the help of purified pirones obtained by the group, it was possible to compare their retention times with the data obtained from leaf extracts of the Cryptocarya species, helping to identify the substances present. Besides the styrylpyrones, the alkaloids menisperin and xantoplanin were also identified in the three species studied. There is a great difference in the amount of... / Mestre

Rhizophytic algae are large, abundant primary producers throughout tropical and subtropical areas worldwide where they grow as an understory in seagrass beds, as well as form mixed or monospecific beds of exclusively rhizophytic algal species. In this dissertation, "rhizophytic algae" refers to coenocytic green algae (Chlorophyta) in the order Bryopsidales that use a net of rhizoids to anchor in unconsolidated sediments. In the development of seagrass beds, rhizophytic algae colonize bare patches and are thought to facilitate seagrass colonization by stabilizing sediments and providing organic matter. However, despite their prominence little is known about many aspects of the ecology of rhizophytic algae. Detailed information on the abundance and biomass of rhizophytic algae at the species level is scarce and the belowground components are seldom quantified. Moreover, rhizophytic algal communities located along the central west coast of Florida have received very little study. At three shallow coastal sites in the Lower Florida Keys and one on the central west coast of Florida, I measured the abundance, biomass, organic content, and morphometric features of the above- and belowground portions of all rhizophytic algal species present along transects in seagrass-algal bed habitat. Relatively diverse assemblages of these algae were present both in areas with and without a seagrass canopy, though dense (greater than or equal to 50%) seagrass cover correlated with decreased algal richness. Rhizophytic algal densities at Keys sites ranged from 68 - 143 thalli m-2 with total dry weights of 76.4 - 226.7 g m-2 with only calcified species present. The west coast of Florida site had the highest aboveground organic biomass (180 g m-2), the highest abundance of rhizophytic algae (365 thalli m-2), and abundant uncalcifed algae of the genus Caulerpa. Morphometric characteristics varied within a species among sites and may reflect differences in abiotic variables such as sediment grain size. The anchoring structures of these algae, made up of fine rhizoids and attached sediment, occupied up to 5.3% of the total volume of the top 5 cm of substrate. My results indicate that across rhizophytic algal species, even within a genus, the production of belowground structure and potential influence on ecosystem function is highly variable and not necessarily related to the aboveground biomass. These results provide new information on belowground structure provided by rhizophytic algal species and characterize the rhizophytic algal community on the central west coast of Florida. The role of rhizophytic algae in seagrass bed succession has been recognized, but little is known about the rate and species composition of colonization of recently created bare patches. In a series of field experiments at three sites on the central west coast of Florida, recruitment by rhizophytic algae into created cleared areas was rapid and dominated by two species of Penicillus and Udotea flabellum. In three weeks, rhizophytic algae were able to recruit, grow to their full height, and bind sufficient sediment to create full-sized holdfasts. Additional field experiments described here show thalli of all of the rhizophytic algal species tested (three species in three genera) were able to regenerate from holdfasts (with small stubs of stipe attached) in a matter of weeks. Overall, my results suggest that belowground structures play a key role in recolonization by, and recovery of, rhizophytic algae after disturbance and are likely important to the long-term persistence of these algal populations. Bryopsidalean algae often have high concentrations of defensive compounds inside their thalli and these terpenoid secondary metabolites possess anti-fouling capability in laboratory tests. Because fouling is ubiquitous in marine environments and epibonts have harmful effects on their hosts, researchers have proposed that rhizophytic algae use these compounds to prevent fouling. For this to be an effective strategy, the compounds must be presented to potential colonizers on the external aboveground surfaces. Thus, I examined the chemistry of rhizophytic algal surfaces using extractions that avoid mechanical damage. Secondary metabolites were not detected in the surface extracts of four species while these compounds were detected in the whole plant extracts. My results, coupled with previous studies on the degradation of these metabolites in seawater and the presence of fouled plants in the field, and suggest non-polar secondary metabolites are not deployed onto the surfaces of rhizophytic algae as a defense against fouling.

Submitted by Victor Alexandre Zonaro (victor.zonaro@gmail.com) on 2017-01-10T14:22:50Z No. of bitstreams: 1 Dissertação - Victor Alexadre Zonaro.pdf: 4719611 bytes, checksum: 075493217f07f41d58c2e71196b60eea (MD5) / Approved for entry into archive by LUIZA DE MENEZES ROMANETTO (luizamenezes@reitoria.unesp.br) on 2017-01-13T17:16:22Z (GMT) No. of bitstreams: 1 zonaro_va_me_araiq.pdf: 4719611 bytes, checksum: 075493217f07f41d58c2e71196b60eea (MD5) / Made available in DSpace on 2017-01-13T17:16:22Z (GMT). No. of bitstreams: 1 zonaro_va_me_araiq.pdf: 4719611 bytes, checksum: 075493217f07f41d58c2e71196b60eea (MD5) Previous issue date: 2016-12-20 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / As 5,6-diidro-2-pironas 6-substituídas, também conhecidas como estirilpironas, são uma importante classe de metabólitos secundários presentes em plantas. São substâncias biologicamente ativas, apresentando como, por exemplo, atividade anticâncer, antioxidante, antifúngica e antiviral. O gênero Cryptocarya, pertence à família Lauraceae e apresenta diversas estirilpironas em suas mais diversas partes: folhas, cascas, sementes e raízes. Foram selecionadas para análise as espécies C. mandioccana, C. moschata e C. botelhensis. Foram utilizadas apenas as folhas, por apresentarem facilidade para coleta e abundância. Este trabalho tem como objetivo a identificação das estirilpironas presentes em três espécies brasileiras de Cryptocarya com o uso das técnicas HPLC-DAD-MS. Foi desenvolvido método cromatográfico para análise do extrato hidroalcoólico das folhas de espécies de Cryptocarya por HPLC-DAD-MS utilizando etanol como fase orgânica na fase móvel. Com os espectros no UV foi possível identificar que as classes de metabólitos presentes nas amostras eram alcaloides, flavonoides e estirilpironas. Utilizando os dados de MS e MS/MS, foi possível a caracterização de estirilpironas presentes nos extratos, assim como a sugestão de suas fragmentações por espectrometria de massas, além da identificação dos íons m/z 163 e m/z 189 característicos da fragmentação das estirilpironas. Foi detectado o íon m/z 423 no extrato de C. moschata, referente a um possível dímero da goniotalamina, sem relato anterior para as espécies de Cryptocarya. Com o auxilio de pironas purificadas obtidas pelo grupo, foi possível a comparação de seus tempos de retenção com os dados obtidos a partir dos extratos de folhas das espécies de Cryptocarya ajudando na identificação das substâncias presentes. Além das estirilpironas, foram identificados os alcalóides menisperina e xantoplanina nas três espécies estudadas. Nota-se uma grande diferença na quantidade de metabólitos entre as espécies analisadas, sendo que a C. mandioccana é a mais rica em estirilpironas. / The 6-substituted 5,6-dihydro-2-pyrones, also known as styrylpyrones, are an important class of secondary metabolites present in plants. They are biologically active substances, presenting, for example, anticancer, antioxidant, antifungal and antiviral activity. The genus Cryptocarya belongs to the family Lauraceae and presents several styrylpyrones in its most diverse parts: leaves, barks, seeds and roots. The species C. mandioccana, C. moschata and C. botelhensiswere selected for analysis. We chose to use only leaves, because they are easy to collect and abundant. The objective of this work is to identify the styrylpyrones present in three Brazilian Cryptocarya species using the HPLC-DAD-MS techniques. A chromatographic method was developed to analyze the hydroalcoholic extract from leavesCryptocarya species by HPLC-DAD-MS using ethanol as the organic solvent in the mobile phase. With the UV spectra were possible to identify the classes of metabolites present in the samples as alkaloids, flavonoids and styrylpyrones. Addtitionaly, with the MS and MS2 data, was possible to characterize the styrylpyrones present in the extracts, as well as the suggestion of their fragments by mass spectrometry, in addition to the identification of íons m/z 163 and m/z 189 characteristics of the fragmentation of styrylpirones. The m/z 423 ion detected in C. moschata extract was tentatively attributed to a goniotalamine dimer, with no previuous reports for Cryptocarya species. With the help of purified pirones obtained by the group, it was possible to compare their retention times with the data obtained from leaf extracts of the Cryptocarya species, helping to identify the substances present. Besides the styrylpyrones, the alkaloids menisperin and xantoplanin were also identified in the three species studied. There is a great difference in the amount of metabolites among the analyzed species, with C. mandioccana being the richest in styrylpyrones.

Submitted by (edna.saturno@ufrpe.br) on 2016-06-29T14:39:18Z No. of bitstreams: 1 Milena Dutra da Silva.pdf: 584735 bytes, checksum: 4ed04e843193f8526a523d7cedcbdb96 (MD5) / Made available in DSpace on 2016-06-29T14:39:18Z (GMT). No. of bitstreams: 1 Milena Dutra da Silva.pdf: 584735 bytes, checksum: 4ed04e843193f8526a523d7cedcbdb96 (MD5) Previous issue date: 2008-02-04 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Myracrodruon urundeuva Allemão (Anacardiaceae), Sideroxylon obtusifolium (Roem. & Schult.) T. D. Penn. (Sapotaceae) and Zizyphus joazeiro Mart. (Rhamnaceae) are native species of the caatinga, widely used as medicinal, specially parts from the stem bark of adult individuals. In general, it is collected hazardously and can let the plant to die and, consequently, reduce the biodiversity. This study aimed to characterize the anatomical, histochemical and phytochemical profile of these species, to the botanical certification, to identify and to localize the accumulation of the metabolites, comparing young stem and mature leaves in young and adult individuals. Usual methods in plant anatomy, phytochemical tests to detect the metabolites classes and specific phytochemical tests to phenolic compounds, iridoids and starch were made. M. urundeuva shows amphystomatic leaves with anomocytic stomata; unicellular simple and glandular trichomes, more abundant over the veins; idioblasts with prismatic crystals in the spongy parenchyma; secretory ducts associated with the phloem in the main vein of the leaf vein, in the petiole and in the stem. S. obtusifolium shows hypostomatic leaves with actinocytic stomata; tector trichomes in the leaf lamina and in the petiole; unistratified hypodermis, with idioblasts with druses; sclereids in the mesophyll; secretory ducts in the medulla of the petiole and the stem. Z. joazeiro shows anomocytic and tetracytic stomata, unistratified hypodermis, with idioblasts with druses, braciform cells in the spongy parenchyma, prismatic crystals in the main vein of the leaf, cap of gelatinous fibers in the stem. Z. joazeiro shows cumarin molecules and cinamic derivates in the stem, only in adult individuals. The other species show similar characteristics related to the metabolites in the stem and in the leaves, in young and adult individuals with different stadium of development. / Myracrodruon urundeuva Allemão (Anacardiaceae), Sideroxylon obtusifolium (Roem. & Schult.) T. D. Penn. (Sapotaceae) e Zizyphus joazeiro Mart. (Rhamnaceae) são espécies nativas da caatinga, amplamente utilizadas como medicinais, especialmente de partes do caule de plantas adultas. Em sua grande maioria, eles são coletados de forma danosa, podendo ocasionar a morte do vegetal e, por conseguinte, a perda da biodiversidade. Este estudo objetivou caracterizar o perfil anatômico, histoquímico e fitoquímico dessas espécies, para certificação botânica, identificação e localização de acúmulo de metabólitos, comparando partes jovens de caule e folhas maduras em indivíduos jovens e adultos. Foram utilizados métodos usuais em anatomia vegetal, testes fitoquímicos para detecção das classes de metabólitos e testes histoquímicos específicos para compostos fenólicos, triterpenos e amido. M. urundeuva apresenta folhas anfiestomáticas com estômatos anomocíticos; tricomas unicelulares simples e glandulares, em maior quantidade sobre as nervuras; idioblastos com cristais prismáticos no parênquima esponjoso; ductos secretores associados ao floema na nervura principal da lâmina foliar, no pecíolo e no caule. S. obtusifolium apresenta folhas hipoestomáticas com estômatos actinocíticos; tricomas tectores na lâmina foliar e no pecíolo; hipoderme uniestratificada, com idioblastos contendo drusas; esclereídeos no mesofilo; ductos de secreção na região medular do pecíolo e do caule. Z. joazeiro apresenta estômatos anomocíticos e tetracíticos, hipoderme uniestratificada, com idioblastos contendo drusas, células do esponjosobraciformes, cristais prismáticos na nervura principal da folha, calotas de fibras gelatinosas no caule. Z. joazeiro apresentou moléculas de cumarinas e derivados cinâmicos no caule, apenas na planta adulta, as demais espécies apresentaram características comuns quanto à presença dos metabólitos e sua localização no caule e nas folhas, em indivíduos jovens e adultos. Isto indica que os metabólitos ocorrem em indivíduos com graus de desenvolvimento variado.

Submitted by Fabio Sobreira Campos da Costa (fabio.sobreira@ufpe.br) on 2016-07-22T12:40:38Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Dissertação - Nataliane Marques de Vasconcelos.pdf: 1186432 bytes, checksum: 7aaaefe15061c83ecc86656db0d731f1 (MD5) / Made available in DSpace on 2016-07-22T12:40:38Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Dissertação - Nataliane Marques de Vasconcelos.pdf: 1186432 bytes, checksum: 7aaaefe15061c83ecc86656db0d731f1 (MD5) Previous issue date: 2016-02-26 / CNPq / A resistência microbiológica aos antibióticos constitui uma série problemas de saúde pública por dificultar o tratamento das infecções. As actinobactérias são fontes importantes para a descobertas de novas moléculas com atividades biológicas. A este grupo, o gênero Streptomyces possuem dentre outros, dois grupos de enzimas multimoduladoras conhecidas como policetídeo sintase (PKS) e peptídeo sintase não ribossomal (NRPS), genes relacionados com a produção de metabólitos secundários. O presente trabalho teve como objetivo investigar o potencial in vitro de metabólitos bioativos produzidos por Actinobactérias isoladas do bioma Caatinga com atividade contra diferentes isolados clínicos de Candida spp. Após ensaio primário das 45 actinobactérias apenas a linhagem PR- 32 apresentou atividade contra Candida spp, com halos de até 20 mm no meio ISP2. Posteriormente, essa linhagem foi cultivada em seis diferentes meios de cultura sendo observada melhor produção do metabólito secundário no meio 400 em 48 horas (h) de fermentação. A determinação da concentração mínima inibitória (CMI) foi determinada a partir do extrato etanólico da biomassa de PR- 32 em pH 7.0 e foi evidenciada uma CMI entre 31,25 μg/mL a 3,9 μg/mL para as leveduras testadas. A cinética de morte reforçou o resultado da CMI e mostrou que no período de 4-8 h o extrato inibiu as cepas de Candida spp. A caracterização da actinobactéria foi identificada por metodologias clássicas e pela pesquisa do gene 16S rRNA como Streptomyces sp. PR- 32. Os resultados desta caracterização sugerem uma possível espécie nova, contudo outras análises ainda precisam ser realizadas. Foi evidenciada a presença do gene nrps com aproximadamente 750 kb. Diante destes resultados podemos concluir que Streptomyces sp. PR- 32 é um isolado promissor para produção de compostos antifúngicos, sendo possível sugerir que a atividade biológica deste metabólito secundário é regulado por peptídeo sintase não ribossomal (NRPS). / The microbial resistance to antibiotics is a series of public health problems for hindering the treatment of infections. The actinomycetes are important sources for new molecules with biological activities discovered. In this group, the genus Streptomyces have among others, multimoduladoras two groups of enzymes known as polyketide synthase (PKS) and non-ribosomal peptide synthase (NRPS), genes involved in production of secondary metabolites. This study aimed to investigate the potential in vitro bioactive metabolites produced by isolated Actinobacteria biome Caatinga with activity against different clinical isolates of Candida spp. After screening of actinomycetes in 45 primary test only the PR- 32 strain showed activity against Candida spp, with halos of up to 20 mm in the middle ISP2. Subsequently, this strain was grown in six different culture media is best seen in secondary metabolite production means 400 at 48 h of fermentation. The determination of the minimum inhibitory concentration (MIC) was determined from the ethanolic extract of the biomass of PR- 32 at pH 7.0 and one MIC was observed between 31.25 mg / mL 3.9 mg / mL for yeast tested. The kinetics of death reinforced the result of CMI and showed that in the 4-8 hour period the extract inhibited the strains of Candida spp. The characterization of actinobacteria was identified by classical methods and research 16S rRNA gene as Streptomyces sp. PR- 32. The results of this characterization suggests a possible new species, but other tests that must be performed. the presence of the NRPS gene of approximately 750 kb was observed. From these results we conclude that Streptomyces sp. PR- 32 is a promising isolated to produce antifungal compounds, it is possible to suggest that the biological activity of this secondary metabolite is regulated by peptide synthase not ribosomal (NRPS).

Esponjas do gênero Dysidea (Ordem: Dictyoceratida) caracterizam-se por apresentarem grande diversidade de metabólitos secundários, muitos dos quais apresentam potentes atividades biológicas. Este trabalho descreve o estudo de duas amostras de esponjas da espécie Dysidea robusta, DR1 e DR2, coletadas no litoral da Bahia em 1999. Tal estudo consistiu no fracionamento das amostras, nas análises de seus extratos brutos por LC-MS e técnicas de RMN- mono e bidimensionais. Dentre os extratos de DR1, a fração DR1-EP-5A obtida do extrato éter de petróleo apresentou uma mistura de três ceramidas saturadas (22, 23 e 24). Já da amostra DR2, as frações do extrato aquoso DR2-AQ-6B e DR2-AQ-6D mostraram ser constituídas por derivados do ácido pirodisinóico (18, 19, 20 e 21). Com exceção do ácido pirodisinóico (18), os demais compostos isolados ainda não foram relatados na literatura. / Sponges of the genus Dysidea (Order: Dyctioceratida) are characterized as sources of several biologically active secondary metabolites. This work describes the study of two samples of D.robusta, DR1 and DR2, both collected at the Bahia state coastline, in 1999. The investigation aimed the crude extract fractionation and analysis by LC-MS and by 1D and 2D NMR techniques. Among the extracts DR1, the fraction DR1-EP-5A obtained from the petroleum ether extract showed a mixture of three saturated ceramides, represented by 22, 23 and 24. From the DR2 sample, the fractions obtained from the aqueous extract DR2-AQ-6B and -6D presented pirodisinoic acid derivates 18, 19, 20 and 21. Except for pyrodisinoic acid (18), all other isolated compounds haven´t been reported in the literature yet.

Plant cell culture is an attractive platform technology for production and supply of important plant derived medicinals. A unique characteristic of plant cells is the ability to grow as multicellular aggregates in suspension. The presence of these non-uniform aggregates results in creation of distinct microenvironments, which can induce variations in cellular metabolism (e.g., growth, oxygen consumption and secondary metabolite synthesis). This heterogeneity can lead to unpredictable and suboptimal performance in large scale bioprocesses. One example is the Taxus cell culture system, which produces a widely used chemotherapeutic drug - paclitaxel (Taxol ®). Despite extensive process engineering efforts which have led to increased yields of paclitaxel, Taxus cells exhibit variability in productivity that is poorly understood. Elicitation of Taxus cultures with methyl jasmonate (MeJA) induces the accumulation of paclitaxel, but to varying extents in culture. A significant negative correlation was observed between paclitaxel level and mean aggregate size of the culture, demonstrating the relevance of measuring, and potentially controlling aggregate size during long term subculture. Understanding the regulation of gene expression can provide rational engineering strategies to control variability and optimize performance of Taxus cell cultures. Biosynthetic pathway gene analyses revealed upregulation of genes upon elicitation with MeJA; results also suggested additional molecular regulatory points outside of the biosynthetic pathway. In order to fully understand Taxus molecular regulation and the relationship to paclitaxel production variability, a transcriptome-wide analysis using next generation sequencing (454 and Illumina) methods was performed. Several pathways outside of paclitaxel biosynthesis were found active upon MeJA elicitation. Global comparison of gene expression amongst cultures accumulating different levels of paclitaxel is being performed to completely understand the interactions amongst the paclitaxel biosynthetic pathway and other complimentary and competing pathways to suggest effective targets for metabolic engineering. This work collectively represents the first molecular studies to understand metabolic regulation in Taxus cell cultures. Apart from inducing paclitaxel biosynthesis, MeJA decreases cell growth in Taxus cell cultures. The MeJA-mediated repression of cell growth was shown to correlate with inhibition of cell cycle progression as evident both at the culture level through flow cytometric analyses and at the transcriptional level by repression of key cell cycle-associated genes. Results from this study provide valuable insight into the mechanisms governing MeJA perception and subsequent events leading to repression of Taxus cell growth.

The fungus Macrophomina phaseolina (Tassi) Goidanich (Mp) is the causal agent of charcoal rot in soybean and infects over 500 plant species worldwide. Mp produces various mycotoxins and is suspected of utilizing a toxin-mediated process to penetrate host tissue. Identification and evaluation of secondary metabolites produced by Mp will further elucidate the pathogenesis mechanisms used by the fungus. Mp cultures isolated from soybean were evaluated for phytotoxicity in a hydroponic soybean bioassay and chemically analyzed by LC-MS/MS. All Mp cultures at two dilutions induced phytotoxicity symptoms including chlorosis, necrosis, wilting, stunting, and death. Analysis identified 13 unreported secondary metabolites including mellein, a compound with various biological activities. The phytotoxicity of mellein was evaluated against soybean seedlings in hydroponic culture, and symptoms of wilting and stunting were observed at levels above 40 MUg/L. Observations suggest that mellein does not directly contribute to the phytotoxic effects of Mp cultures.

No description available.